ISHS Acta Horticulturae 812: III International Symposium on Acclimatization and Establishment of Micropropagated Plants
ASYMBIOTIC CULTIVATION IN VITRO OF THE ENDANGERED ORCHID CYPRIPEDIUM CALCEOLUS L. AND SOME ASPECTS OF EX VITRO GROWTH |
| Authors: | D. Klavina, I. Druva-Lusite, A. Gailite |
| Keywords: | aseptic culture, cultivation, germination, lady's slipper orchid, orchid mycorrhiza, organogenesis |
| Abstract:
Ex situ conservation of endangered plant species is one of the most important objectives in botanic gardens. Micropropagation is an appropriate method for complex wild plant conservation systems. There are some difficulties in germination and growth of various orchid species, therefore in vitro culture experiments were performed with Cypripedium calceolus L. Seeds collected from natural habitats were germinated asymbiotically in the dark on 2 different media, each without and with 6-benzylaminopurine (BA). Seed germination was significantly influenced by medium composition as well as BA incorporation in medium. BA addition enhanced germination in both media and the best germination was achieved on half diluted Norstog medium with BA. Germinated seeds went through the embryo or protocorm stage, protocorm growth stage (without production of rhizoids in our conditions), bud development, abundant development of roots, enlargement of leaf scale and its opening and production of greenish leaves. Seedling proliferation and cluster production were observed and shoot development was sometimes difficult to obtain. BA, kinetin or zeatin induced some increase in seedling root and shoot number within 2 months, with zeatin being the most efficient. However, differences were minimal and using uneven seedling material may have influenced these results. A 2-3 months storage period at 5°C stimulated establishment of seedlings ex vitro. After 2 months ex vitro, the root mycorrhization percentage was between 1.0-8.9%. | |
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